Construction Fermentation of Engineered Strain and Properties of Recombinant ?-glutamyltranspeptidase
- VernacularTitle:?-谷氨酰基转肽酶(GGT)基因工程菌的构建及其发酵条件的初步研究
- Author:
Na WANG
;
Jie ZHANG
;
Wei SHEN
;
Xue-Ming TANG
;
Ge-Jian ZHU
;
- Publication Type:Journal Article
- Keywords:
?-Glutamyltranspeptidase Theanine Biotransformation Fermentation condition Plasmid stability
- From:
China Biotechnology
2006;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
The ?glutamyltranspeptidase encoding gene(ggt) from Bacillus subtilis SYU 20016 was amplified by PCR. The ggt gene was inserted in pBV220 to yield the recombinant expression vector pBV220ggt. Overexpression of ggt in E.coli JM109 was achieved with pBV220ggt. SDSPAGE analysis showed an overexpressed recombinant product at about 65kDa,consistent with the molecular weight predicted from gene sequence. The ferment conditions of r-glutamyltranspeptidase were also discussed. The optimum temperature and pH for the enzyme were determined as 30℃ and 7.2 respectively.The cultures were incubated at 42℃ for 4h with broth volume 20ml/250ml flask and the yield of 6U/ml was obtained, enzyme activity of B. subtilis NX2 was only 3.2 U/ml.
